Clin Immunol, 1999 Feb, 90(2), 256 - 65 Involvement of nitric oxide in protecting mechanism during experimental cryptococcosis; Rossi GR et al.; In the present study we investigated the role of nitric oxide (NO) in the effector mechanisms of host defense against Cryptococcus neoformans in vivo . Our results showed an increase of NO produced by the peritoneal macrophages from 14-days infected rats compared with normal rats . These cells were capable of killing C . neoformans to a greater extent than macrophages from noninfected rats (80% vs 20%, respectively) . The killing of C . neoformans by infected cells was efficiently inhibited (80% to 35%, P < 0.001) by adding aminoguanidine (AG) to the cultures . We observed that in vivo administration of AG to the infected animals efficiently inhibited the metabolism producing NO and failed to affect that of normal animals . When the NO synthase (NOS) was inhibited in vivo in the infected animals, a marked increase of the fungi charge in the organs was observed with respect to the normal animals treated with AG . We also observed that the course of the infection is drastically modified after the inhibition of NO production because all the animals infected and treated with AG died from cryptococcosis before 20 days postinfection (p.i.) . These results indicate that NO is a crucial molecule in the effector mechanisms in this infection model . J Clin Oncol, 1999 Feb, 17(2), 554 - 60 Value of combined approach with thallium-201 single-photon emission computed tomography and Epstein-Barr virus DNA polymerase chain reaction in CSF for the diagnosis of AIDS-related primary CNS lymphoma; Antinori A et al.; PURPOSE: To determine the diagnostic capability of thallium-201 (201Tl) single-photon emission computed tomography (SPECT) combined with Epstein-Barr virus DNA (EBV-DNA) in CSF for the diagnosis of AIDS-related primary CNS lymphoma (PCNSL) . PATIENTS AND METHODS: All human immunodeficiency virus (HIV)-infected patients with focal brain lesions observed between June 1996 and March 1998 underwent lumbar puncture and 201Tl SPECT . Each CSF sample was tested with polymerase chain reaction (PCR) for EBV-DNA . RESULTS: Thirty-one patients were included, 13 with PCNSL and 18 with nontumor disorders . In 11 PCNSL patients, EBV-DNA was positive . Thallium-201 uptake ranged from 1.90 to 4.07 in PCNSL cases (mean, 2.77; 95% confidence interval {CI}, 2.35 to 3.19) and from 0.91 to 3.38 in nontumor patients (mean, 1.62; 95% CI, 1.30 to 1.94) (P<.0002) . Using a lesion/background ratio of 1.95 as cutoff, a negative SPECT was found in one PCNSL case and 16 nonneoplastic cases . A cryptococcoma and a tuberculoma showed highly increased 201Tl uptake . Epstein-Barr virus DNA was never detected in nonneoplastic patients . For PCNSL diagnosis, hyperactive lesions showed 92% sensitivity and 94% negative predictive value (NPV), whereas positive EBV-DNA had 100% specificity and 100% positive predictive value . The presence of increased uptake and/or positive EBV-DNA had 100% sensitivity and 100% NPV . CONCLUSION: Combined SPECT and EBV-DNA showed a very high diagnostic accuracy for AIDS-related PCNSL . Because PCNSL likelihood is extremely high in patients with hyperactive lesions and positive EBV-DNA, brain biopsy could be avoided, and patients could promptly undergo radiotherapy or multimodal therapy . On the contrary, in patients showing hypoactive lesions with negative EBV-DNA, empiric anti-Toxoplasma therapy is indicated . In patients with discordant SPECT/PCR results, brain biopsy seems to be advisable. Rinsho Shinkeigaku, 1998 Sep, 38(9), 831 - 7 {Serial MRI findings in patients with CNS cryptococcosis}; Kumazawa K et al.; We reported the serial magnetic resonance imaging (MRI) findings of two patients with central nervous system (CNS) cryptococcal infection without AIDS . The diagnosis of CNS cryptococcosis was made by visualizing the fungi in the CSF with the India ink test, detecting cryptococcal antigens, and culturing the fungus . Both patients had dilated perivascular Virchow-Robin (V-R) spaces, which were defined as small rounded lesions greater less than 3mm diameter that were hyperintense on T2-weighted images . They were present in the basal ganglia, brainstem and cerebral white matter . Case 1 had bilateral parietal arachnoid cyst which was thought to represent a focal collection of organisms and mucoid material within subarachnoid space . Abnormal optochiasmatic arachnoid enhancement detected in case 2, who had complete loss of vision . With disease progression perivascular V-R increased in size, resulting in the developing cryptococomas which were defined as rounded lesions greater than 3mm diameter, and were hyperintense on T2-weighted images in the basal ganglia, cerebellum and cerebral white matter . In follow-up MRI of those patients, radiological progression was seen despite appropriate treatment and falling CSF cryptococcal antigens . In conclusion, this spectrum of MRI appearances in CNS cryptococcosis reflects the pathological mechanism of invasion by the fungus, and may be relatively specific for cryptococcosis. Appl Microbiol Biotechnol, 1999 Jan, 51(1), 40 - 5 Production of sophorolipids from whey: development of a two-stage process with Cryptococcus curvatus ATCC 20509 and Candida bombicola ATCC 22214 using deproteinized whey concentrates as substrates; Daniel HJ et al.; In order to produce sophorolipids from whey, thereby lowering the lactose content and biological oxygen demand, a two-step batch cultivation process was developed including medium sterilization by filtration . In the first step, whey was sterilized by a combination of crossflow and sterile filtration . Because the sophorolipid-producing yeast Candida bombicola ATCC 22214 was not able to use lactose as a carbon source directly, the oleaginous yeast Cryptococcus curvatus ATCC 20509 was grown on deproteinized whey concentrates (DWC) . With 1:1 diluted DWC-20, lactose was consumed as the carbon source and biomass (24 g/l dry weight content) as well as single-cell oil (SCO, 10 g/l) were produced . The cultivation broth was disrupted with a glass bead mill and it served as medium for growth (29 g cell dry mass/l) and sophorolipid production (12 g/l) of the yeast C . bombicola. Med Mycol, 1998 Oct, 36(5), 341 - 4 First isolation of Cryptococcus neoformans var . gattii, serotype C, from the environment in Colombia; Callejas A et al.; The natural habitat of Cryptococcus neoformans var . gattii, serotype B in the environment was established by Australian investigators who demonstrated its association with species of Eucalyptus . The aim of the present study was to search for the habitat of this variety in a city of Colombia, where clinical cases due to this variety occur with great frequency . For a period of 5 months detritus, vegetable material and air samples in and around 68 almond trees (Terminalia catappa) located in the city were studied . C . neoformans var . gattii serotype C was the only variety isolated from two of the 68 trees sampled . These trees were positive for 4 of the 5 months during which they were studied . From the first positive sample kept under refrigeration, it was possible to isolate the fungus up to 3 months later . This is the first report of the isolation of serotype C from the environment . More studies are required in order to establish the ecological significance of this finding. Med Mycol, 1998 Oct, 36(5), 335 - 9 First report of Cryptococcus laurentii meningitis and a fatal case of Cryptococcus albidus cryptococcaemia in AIDS patients; Kordossis T et al.; We report the first case of Cryptococcus laurentii meningitis and a rare case of Cryptococcus albidus cryptococcaemia in AIDS patients . Both infections were treated with amphotericin B and flucytosine . The C . laurentii meningitis was controlled after 2 weeks of treatment with no evidence of infection 20 months later . The patient with C . albidus cryptococcaemia, despite the amphotericin B/flucytosine combination therapy, died on the 14th day of treatment . The minimum inhibitory concentrations (MICs) for C . laurentii, as determined by Etest on RPMI 1640 agar, were 0.25 microg ml(-1) of amphotericin B, 1.25 microg ml(-1) flucytosine, 4 microg ml(-1) fluconazole, 0.50 microg ml(-1) itraconazole and 1.0 microg ml(-1) of ketoconazole . The MIC of amphotericin B for C . albidus was 0.5 microg ml(-1), flucytosine 1.25 microg ml(-1), fluzonazole 4 microg ml(-1), itraconazole 0.5 microg ml(-1) and ketonazole 0.25 microg ml(-1) . The agreement of the amphotericin B MIC values obtained in antibiotic medium 3 by the broth microdilution method, with those obtained on casitone medium by Etest, was within a two-dilution range for both isolates . C . laurentii may cause meningitis and may also involve the lungs in AIDS patients. Med Mycol, 1998 Oct, 36(5), 305 - 11 Cryptococcus neoformans isolated from human dwellings in Rio de Janeiro, Brazil: an analysis of the domestic environment of AIDS patients with and without cryptococcosis; Passoni LF et al.; One hundred and fifty-four human dwellings in the metropolitan area of Rio de Janeiro, Brazil were studied . A total of 824 samples of indoor dust, outdoor soil and avian droppings were collected . Cryptococcus neoformans var . neoformans was isolated from 20 (13%) dwellings, comprising five (15.6%) of 32 dwellings of patients with AIDS-associated cryptococcosis; four (8.9%) of 45 dwellings of patients with AIDS but without cryptococcosis; and 11 (14.3%) of 77 dwellings of apparently healthy individuals (P>0.05) . The principal factor associated with domiciliar contamination by C . neoformans var . neoformans was the presence of avians in the domestic environment or nearby the home . Cryptococcosis was more frequent among AIDS patients residing in dwellings from which C . neoformans var . neoformans was isolated than among AIDS patients from whose domestic environment the fungus was not demonstrated by the methods used (odds ratio (OR)=2.05) . These findings suggest that the distribution of C . neoformans var . neoformans in Rio de Janeiro is not restricted to the classically known biotopes as well as reinforce the possibility of exogenous infection in opportunistic cryptococcosis, including exogenous infection acquired in the domestic environment. Int J Antimicrob Agents, 1999 Jan, 11(1), 65 - 8 Characterization of antimicrobial agents extracted from Asterina pectinifera; Choi DH et al.; The extracts from the starfish Asterina pectinifera obtained using various organic solvents and distilled water were tested for their antimicrobial activities against fungi and yeasts . The methanol and water extracts were found to be the most active and Aspergillus spp . and Cryptococcus neoformans proved to be sensitive species . The extracts are polar, stable to a wide range of pH and a high temperature, and possess a very distinctive UV spectrum . The colour reactions for functional groups of antimicrobial substances indicate the presence of some distinctive chemical properties common to steroidal oligoglycosides which results suggest that the antimicrobial activity is due to structures common to steroidal oligoglycosides. J Clin Microbiol, 1999 Apr, 37(4), 1170 - 2 Serotyping of Cryptococcus neoformans isolates from clinical and environmental sources in Spain; Baro T et al.; We determined biovars and serotypes of 154 isolates of Cryptococcus neoformans from clinical and environmental sources from different areas of Spain . All clinical isolates belonged to C . neoformans var . neoformans . Serotypes showed an irregular distribution . C . neoformans var . gattii serotype B was isolated from necropsy specimens from goats with pulmonary disease. J Clin Microbiol, 1999 Apr, 37(4), 981 - 6 Detection of cell wall mannoprotein Mp1p in culture supernatants of Penicillium marneffei and in sera of penicilliosis patients; Cao L et al.; Mannoproteins are important and abundant structural components of fungal cell walls . The MP1 gene encodes a cell wall mannoprotein of the pathogenic fungus Penicillium marneffei . In the present study, we show that Mp1p is secreted into the cell culture supernatant at a level that can be detected by Western blotting . A sensitive enzyme-linked immunosorbent assay (ELISA) developed with antibodies against Mp1p was capable of detecting this protein from the cell culture supernatant of P . marneffei at 10(4) cells/ml . The anti-Mp1p antibody is specific since it fails to react with any protein-form lysates of Candida albicans, Histoplasma capsulatum, or Cryptococcus neoformans by Western blotting . In addition, this Mp1p antigen-based ELISA is also specific for P . marneffei since the cell culture supernatants of the other three fungi gave negative results . Finally, a clinical evaluation of sera from penicilliosis patients indicates that 17 of 26 (65%) patients are Mp1p antigen test positive . Furthermore, a Mp1p antibody test was performed with these serum specimens . The combined antibody and antigen tests for P . marneffei carry a sensitive of 88% (23 of 26), with a positive predictive value of 100% and a negative predictive value of 96% . The specificities of the tests are high since none of the 85 control sera was positive by either test. Hunan Yi Ke Da Xue Xue Bao, 1997, 22(6), 514 - 6 {Nitric oxide, TNF-alpha and IL-8 in cerebrospinal fluids of tuberculous and cryptococcic meningitis}; Chen L et al.; The contents of NO, TNF-alpha and IL-8 in the cerebrospinal fluids (CSF) of patients with tuberculous and cryptococcic meningitis were detected . The results showed that the concentration of NO2-/NO3- and the levels of TNF-alpha and IL-8 in CSF of the two kinds of meningitis were higher than those of normal CSF, and the concentration of NO2-/NO3- correlated positively to the content of TNF-alpha . The results indicate that the over production of NO, TNF-alpha and IL-8 in CSF may involve in the tissue inflammation and damage of central nerve system infection. J Lab Clin Med, 1999 Mar, 133(3), 274 - 88 Urokinase is required for T lymphocyte proliferation and activation in vitro; Gyetko MR et al.; We have previously demonstrated that urokinase-deficient (uPA-/-) mice do not increase lung T lymphocyte number and fail to mount protective immune responses during pulmonary Cryptococcus neoformans infection . These observations suggest a previously unconsidered role for urokinase-type plasminogen activator (uPA) in T lymphocyte-mediated immune responses . Accordingly, we sought to determine whether uPA is required for T cell receptor-mediated (TCR-mediated) lymphocyte proliferation and activation . Splenocytes from uPA-/- and uPA+/+ mice were stimulated with concanavalin A (Con A) . The uPA-/- mice had diminished T cell proliferation as compared with uPA+/+ mice . Coculturing uPA-/- T cells with uPA+/+ accessory cells led to the restoration of proliferation . Similarly, T cell proliferation induced by CD3 cross-linking was diminished in uPA-/- mice as compared with uPA+/+ mice . T lymphocyte activation, defined as the induced expression of antigens and the elaboration of cytokines, was determined . The expression of CD69 and that of CD49d were diminished in response to Con A stimulation in uPA-/- mice as compared with uPA+/+ mice . The elaboration of cytokines in response to Con A was also altered in the uPA-/- mice . The production of the Th1 cytokines interferon-gamma and interleukin-12 was diminished in uPA-/- mice as compared with uPA+/+ mice . The uPA-/- mice produced increased amounts of interleukin-10, a Th2 cytokine . We conclude that the lack of uPA results in impaired T cell activation and proliferation in response to TCR-mediated signaling and the expression of a less Th1-polarized profile of cytokines . These findings suggest that the inability of uPA-/- mice to combat Cryptococcus neoformans infection may be caused by the impairment of T lymphocyte immune responses in the absence of uPA. Skeletal Radiol, 1999 Jan, 28(1), 49 - 51 Cryptococcoma of the sacrum; Noh HM et al.; Cryptococcoma of the sacrum was the initial presentation of systemic cryptococcosis in a patient on chronic steroid therapy for autoimmune hepatitis . The bone lesion was the only overt manifestation of systemic cryptococcal disease, which preceded other clinical manifestations and led to the subsequent diagnosis of systemic infection. Clin Diagn Lab Immunol, 1999 Mar, 6(2), 266 - 8 Variants of a Cryptococcus neoformans strain elicit different inflammatory responses in mice; Chen LC et al.; The virulence of Cryptococcus neoformans isolates with high and low extracellular proteolytic activity was investigated in mice . No consistent relationship between proteolytic activity and virulence was observed, but isolates derived from one strain were shown to elicit different inflammatory responses. Curr Opin Microbiol, 1998 Aug, 1(4), 381 - 9 Gene disruption to evaluate the role of fungal candidate virulence genes; Kwon-Chung K; Gene disruption is a powerful genetic tool that can define pathogenic or virulence factors . In the past two years gene disruption approaches have been used to identify fungal virulence genes . The capsule genes, an alpha subunit of G protein and certain kinases of Cryptococcus neoformans have clearly been demonstrated to be associated with pathogenicity . In Candida albicans at least four genes involved in hyphal formation have been disrupted and tested for virulence . In other fungi, such as Histoplasma capsulatum, however, more efficient gene disruption methods need to be developed before such approaches can be regularly used for identifying virulence genes. J Zoo Wildl Med, 1998 Dec, 29(4), 474 - 8 Sporotrichosis in a nine-banded armadillo (Dasypus novemcinctus); Wenker CJ et al.; An adult female nine-banded armadillo (Dasypus novemcinctus) died in the quarantine station of a private Swiss zoo . Multifocal ulcerative skin lesions and multiple hemorrhages in the lungs were found at necropsy . The spleen was enlarged and dark red . Histologically, there was diffuse granulomatous infiltration, including multinucleated giant cells, of the skin lesions, lungs, spleen, liver, heart, and kidneys . Abundant periodic acid-Schiff-positive yeastlike cells were demonstrated intracellularly in giant cells and extracellularly scattered throughout the tissues . Morphology of the cells varied, with some nonbudding cells resembling Cryptococcus neoformans and others resembling Sporothrix schenckii . A diagnosis of sporotrichosis was confirmed by immunofluorescence studies . This is the first report of sporotrichosis in an armadillo in a zoological garden and the third report of sporotrichosis in D . novemcinctus. No To Shinkei, 1999 Jan, 51(1), 69 - 74 {Steroid-responsive diffuse cerebral white matter lesions in a case of intractable fungal meningoencephalitis}; Suzuki Y et al.; We report a case of fungal meningoencephalitis with steroid-responsive diffuse cerebral white matter lesions . A 49-year-old male developed auditory hallucination, confusion and fever, on April, 1994 . He was diagnosed as having cryptococcal meningoencephalitis based on the detection of cryptococcal antigens in the cerebrospinal fluid (CSF) . Intravenous administration of fluconazole resulted in improvement of his neurologic symptoms and CSF findings . For the next seven months, he was treated with oral fluconazole and the neurological status was stable . However, soon after the dose of fluconazole was tapered, he became confused and febrile, which made him admitted to our hospital . Neurological examination on admission showed disturbance of consciousness, disorientation and meningeal irritation . The CSF examination revealed mild pleocytosis (mostly lymphocytes), elevated protein and normal glucose levels, although fungus was not detected . The T2-weighted image of brain MRI demonstrated diffuse hyperintense lesions in the bilateral cerebral white matters . GD-DTPA enhanced MRI showed spotty enhanced lesions in the periventricular white matters . The neurologic symptoms were once relieved after intravenous administration of fluconazole was started, but two months later, he became comatose and needed ventilatory support, despite amphotericine B therapy . Then, a needle brain biopsy targeting the white matter lesion was done . Histopathology of the specimen showed chronic inflammation with granuloma formation and T lymphycyte infiltrate around the small vessels, though fungus was not detected in the tissue . Combined therapy with corticosteroid and antifungal agents remarkably improved the neurological symptoms as well as the MRI findings . In the present case, fungal infection possibly induced an altered immune reactions which resulted in the steroid responsive diffuse cerebral white matter lesions. Nihon Kokyuki Gakkai Zasshi, 1998 Dec, 36(12), 1038 - 42 {Pulmonary cryptococcosis exhibiting diffuse multiple nodular shadows}; Inui N et al.; We report a case of pulmonary cryptococcosis showing diffuse multiple nodular shadows in all lung fields . A 39-year-old woman with no immunological abnormalities was admitted with complaints of cough and sputum . She had experienced measles 4 weeks prior to admission . Chest x-ray films revealed diffuse nodular opacities throughout the lung fields, a finding suggestive of metastatic lung cancer . Detailed examinations, including transbronchial lung biopsy, were not conclusive . A diagnosis of pulmonary cryptococcosis was made on the basis of findings from video-assisted thoracoscopic biopsy . Primary pulmonary cryptococcosis usually appears as a solitary nodule or limited infiltration . Immunologically compromised hosts commonly demonstrate various abnormal shadows, such as the multiple nodular shadows observed in our patient . It has been reported that measles infection can cause temporary immune suppression . Secondary immunodeficiency resulting from the preceding infection with measles could explain the unusual chest x-ray findings in this case. Eur J Immunol, 1999 Feb, 29(2), 643 - 9 Combined effects of IL-12 and IL-18 on the clinical course and local cytokine production in murine pulmonary infection with Cryptococcus neoformans; Qureshi MH et al.; We reported recently that interleukin (IL)-12 and IL-18 synergistically increased the fungicidal activity of mouse peritoneal exudate cells against Cryptococcus neoformans by inducing the production of interferon (IFN)-gamma by natural killer (NK) cells . To confirm these findings in vivo, we examined the effect of combined treatment using these two cytokines on the course of experimentally induced pulmonary and disseminated cryptococcosis in mice . IL-12 and IL-18 were used at subtherapeutic doses (0.005 and 2 microg/mouse/day, respectively) . A single administration of either cytokine was not effective in protecting mice against the infection, while combined treatment significantly prolonged survival time of infected mice and reduced the lung and brain loads of organisms . These protective effects were associated with elevated IFN-gamma and reduced IL-4 levels in bronchoalveolar lavage fluid . Finally, depletion of NK and gammadelta T cells, but not of CD4+ T cells, by administration of specific antibodies, significantly reduced the production of IFN-gamma in lungs by IL-12/IL-18 treatment during the 7 days of infection . Our results demonstrated that IL-12 and IL-18 protected mice against cryptococcal infection in a synergistic manner by enhancing the local production of IFN-gamma by NK and gammadelta T cells in the early phase of infection and by suppressing the production of IL-4 in lungs. Clin Infect Dis, 1999 Feb, 28(2), 309 - 13 Cryptococcosis in children with AIDS; Abadi J et al.; We compiled the clinical and immunologic features of Cryptococcus neoformans infections in human immunodeficiency virus (HIV)-infected children from 1985 to 1996 in a retrospective case series . Thirty cases of cryptococcosis were identified . These children had a median age of 9.8 years, a median CD4+ cell count of 54/microL at the time of diagnosis, and either a culture positive for C . neoformans or cryptococcal antigen in serum or cerebrospinal fluid . Sixty-three percent of the cases occurred in children vertically infected with HIV and in children between 6 and 12 years of age . The clinical and laboratory characteristics of this pediatric cohort were similar to those of adults with AIDS and cryptococcosis . On the basis of a subset of the cases, a 10-year point prevalence of cryptococcosis among children with AIDS of approximately 1% was estimated. Clin Infect Dis, 1999 Feb, 28(2), 291 - 6 A comparison of itraconazole versus fluconazole as maintenance therapy for AIDS-associated cryptococcal meningitis . National Institute of Allergy and Infectious Diseases Mycoses Study Group; Saag MS et al.; This study was designed to compare the effectiveness of fluconazole vs . itraconazole as maintenance therapy for AIDS-associated cryptococcal meningitis . HIV-infected patients who had been successfully treated (achieved negative culture of CSF) for a first episode of cryptococcal meningitis were randomized to receive fluconazole or itraconazole, both at 200 mg/d, for 12 months . The study was stopped prematurely on the recommendation of an independent Data Safety and Monitoring Board . At the time, 13 (23%) of 57 itraconazole recipients had experienced culture-positive relapse, compared with 2 relapses (4%) noted among 51 fluconazole recipients (P = .006) . The factor best associated with relapse was the patient having not received flucytosine during the initial 2 weeks of primary treatment for cryptococcal disease (relative risk = 5.88; 95% confidence interval, 1.27-27.14; P = .04) . Fluconazole remains the treatment of choice for maintenance therapy for AIDS-associated cryptococcal disease . Flucytosine may contribute to the prevention of relapse if used during the first 2 weeks of primary therapy. J Neurol Sci, 1999 Jan 1, 162(1), 20 - 6 Meningitis in a community with a high prevalence of tuberculosis and HIV infection; Silber E et al.; OBJECTIVES: To evaluate the spectrum of aetiologies, and distinguishing clinical and laboratory features, of meningeal infection in a community with a high prevalence of tuberculosis (TB) and HIV infection . SETTING: A hospital serving mineworkers, originating from rural areas of Southern Africa . DESIGN: Prospective cohort of 60 consecutive lumbar punctures (LPs), performed for suspected meningitis . MEASUREMENTS: Clinical history and examination; concurrent cerebrospinal fluid (CSF) and blood samples; mortality status six months after entry to study . RESULTS: 38 of 57 patients (66.7%) were HIV-1 positive, 59.5% of whom had a CD4 count <200 cells/mm3 . Nine patients had tuberculous meningitis (TBM) and two had tuberculomas; four developed disease while on TB therapy . There was one case of multidrug, and two of isoniazid-resistant TBM . There were nine episodes of cryptococcal meningitis (seven patients), nine of aseptic meningitis, two of neurosyphilis and 20 normal LPs, including four with AIDS dementia complex (ADC) . Ten patients with meningococcal infection, part of a larger outbreak, were significantly younger (p=0.004) . All patients with tuberculous, cryptococcal (most immune-suppressed p<0.001) and aseptic meningitis were HIV-1 positive . Within six months, 19 patients had died . Death was associated with HIV positivity (p=0.004), low CD4 count (p<0.001) and a diagnosis of cryptococcal meningitis, CNS TB or ADC . CONCLUSION: HIV has a major impact on the burden of disease and mortality, with a predominance of opportunistic chronic meningitides, despite a meningococcal outbreak, in this community . Of concern is the development of TBM despite therapy, and the emergence of drug-resistant strains. J Antimicrob Chemother, 1998 Dec, 42(6), 779 - 85 In-vivo therapeutic efficacy in experimental murine mycoses of a new formulation of deoxycholate-amphotericin B obtained by mild heating; Petit C et al.; Heat-induced 'superaggregation' of deoxycholate-amphotericin B (AmB-DOC, Fungizone) was shown previously to reduce the in-vitro toxicity of this antifungal agent . We compared AmB-DOC with the formulation obtained by heating the commercial form (Fungizone, Bristol Myers Squibb, Paris, France) for 20 min at 70 degrees C, in the treatment of murine infections . An improvement of antifungal activity was obtained with heated AmB-DOC formulations due to a lower toxicity which allowed the administration of higher drug doses than those achievable with the commercial preparation . Single intravenous injections of heated AmB-DOC solutions were demonstrated to be two-fold less toxic than unheated ones to healthy mice . For mice infected with Candida albicans, the maximum tolerated dose was higher with heated than with unheated AmB-DOC solutions . In the model of murine candidiasis, following a single dose of heated AmB-DOC 0.5 mg/kg, 85% of mice survived for 3 weeks, whereas at this dose the immediate toxicity of the standard formulation in infected mice restricted the therapeutic efficacy to 25% survival . Both formulations were equally effective in increasing the survival time for murine cryptococcal pneumonia and meningoencephalitis . Injection of heated AmB-DOC solutions at a dose two-fold higher than the maximal tolerated dose observed with the unheated preparation (1.2 mg/kg) increased the survival time by a factor of 1.4 in cryptococcal meningoencephalitis . These results indicate that mild heat treatment of AmB-DOC solutions could provide a simple and economical method to improve the therapeutic index of this antifungal agent by reducing its toxicity on mammalian cells. Folia Histochem Cytobiol, 1998, 36(4), 157 - 65 The ovaries of scale insects (Hemiptera, Coccinea) . Morphology and phylogenetic conclusions; Szklarzewicz T; Coccoids (Coccinea, Coccoidea, Coccomorpha, scale insects, scales) are a highly diverse group of ectoparasitic insects . They comprise 2 subgroups: primitive archaeococcoids (= Orthezioidea sensu Koteja) and advanced neococcoids (= Coccoidea sensu Koteja) . The ovaries of coccoids consist of numerous short telotrophic-meroistic ovarioles . The ovarioles of all investigated species share common characters (e.g . the same mechanism of ovariole development, lack of terminal filaments, occurrence of single oocytes in the vitellaria) supporting the concept of monophyletic origin of this group . Despite these characteristics, the ovaries of archaeococcoids and neococcoids differ in the number of germ cells (oocytes + trophocytes) constituting a single ovariole . In primitive families (Ortheziidae, Margarodidae), this number is relatively large (15-58), whereas in advanced ones (Pseudococcidae, Kermesidae, Eriococcidae, Cryptococcidae, Coccidae, Diaspididae) it is small and usually does not exceed 8 . The comparative analysis of the ovary structure in the representatives of Coccinea and closely related Aphidinea (aphids) has revealed that: (1) the organization of archaeococcoid ovaries is more similar to those of aphids than to neococcoids and (2) during the evolution of Coccinea a gradual reduction in the number of germ cells in ovarioles took place. Am J Respir Crit Care Med, 1999 Mar, 159(3), 733 - 40 Human immunodeficiency virus and the outcome of treatment for new and recurrent pulmonary tuberculosis in African patients; Murray J et al.; The purpose of this study was to evaluate the impact of human immunodeficiency virus (HIV) infection on treatment for tuberculosis (TB) . The study population comprised 28,522 black Southern African gold miners . Patients with sputum culture-positive new or recurrent pulmonary TB diagnosed in 1995 were prospectively enrolled in the cohort . Directly observed therapy (DOT) was practiced and outcomes were assessed at 6 mo after treatment was begun . There were 376 cases of TB (incidence 1,318 per 100,000), of which 190 (50%) were HIV positive and 82 (22%) had recurrent TB . There was no association between HIV status and history of previous TB or drug resistance . Neither the treatment interruption rate (2%) nor the rate at which patients transferred out of the treatment program (1.6%) were associated with HIV status . Excluding deaths, cure rates were similar for HIV-positive and HIV-negative patients (89% versus 88%), but significantly lower in those with recurrent than in those with new TB (77% versus 92%) . Mortality was 0.5% in HIV-negative patients versus 13.7% in HIV-positive patients, and in the latter group was associated with CD4(+) lymphocyte depletion . Autopsy examination showed that in HIV-positive patients, early mortality was due to TB whereas late deaths were most commonly due to cryptococcal pneumonia . The study showed that a well-run TB control program can result in acceptable cure rates even in a population with a very high incidence of TB and HIV infection . Particular vigilance is needed for concurrent infections, which may contribute significantly to mortality during treatment of TB in HIV-positive patients. J Antibiot (Tokyo), 1998 Dec, 51(12), 1081 - 6 BE-31405, a new antifungal antibiotic produced by Penicillium minioluteum . I . Description of producing organism, fermentation, isolation, physico-chemical and biological properties; Okada H et al.; A new antifungal antibiotic, BE-31405, was isolated from the culture broth of a fungal strain, Penicillium minioluteum F31405 . BE-31405 was isolated by adsorption on high porous polymer resin (Diaion HP-20), followed by solvent extraction, precipitation and crystallization . BE-31405 showed potent growth inhibitory activity against pathogenic fungal strains such as Candida albicans, Candida glabrata and Cryptococcus neoformans, but did not show cytotoxic activity against mammalian cells such as P388 mouse leukemia . The mechanism studies indicated that BE-31405 inhibited the protein synthesis of C . albicans but not of mammalian cells. Jpn J Thorac Cardiovasc Surg, 1998 Dec, 46(12), 1267 - 74 {Efficacy of F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) scans in diagnosis of pulmonary nodules}; Orino K et al.; To evaluate the value of F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) scans, we performed FDG-PET scans in 23 patients with indeterminate pulmonary nodules less than 3 cm in size and analyzed these scans qualitatively and semiquantitatively . Histologic specimens were obtained by thoracoscopic excisional biopsy in 16 patients, CT-guided needle aspiration cytology in three, and bronchoscopic brushing cytology in four . Pathological diagnoses were lung cancer in 16 patients, benign inflammation in six, and malignant lymphoma in one . Sensitivity, specificity and accuracy of the FDG-PET scans were 88% (15/17), 67% (4/6) and 83% (19/23), respectively . There were two false-positive cases (organizing pneumonia and cryptococcosis) and two false-negative ones (slow-growing adenocarcinoma and malignant lymphoma) . Although a few false-positive cases of granulomatous disease were yielded, the FDG-PET scans were highly sensitive in the detection of lung cancer . We conclude that the FDG-PET scanning in a useful diagnostic imaging modailty in the management of indeterminate pulmonary nodules. Biochim Biophys Acta, 1999 Feb 24, 1453(2), 261 - 72 Interaction of lipopolysaccharide with human small intestinal lamina propria fibroblasts favors neutrophil migration and peripheral blood mononuclear cell adhesion by the production of proinflammatory mediators and adhesion molecules; Chakravortty D et al.; Fibroblasts are important effector cells having a potential role in augmenting the inflammatory responses in various diseases . In infantile diarrhea caused by enteropathogenic Escherichia coli (EPEC), the mechanism of inflammatory reactions at the mucosal site remains unknown . Although the potential involvement of fibroblasts in the pathogenesis of cryptococcus-induced diarrhea in pigs has been suggested, the precise role of lamina propria fibroblasts in the cellular pathogenesis of intestinal infection and inflammation caused by EPEC requires elucidation . Earlier we reported the lipopolysaccharide (LPS)-induced cell proliferation, and collagen synthesis and downregulation of nitric oxide in lamina propria fibroblasts . In this report, we present the profile of cytokines and adhesion molecules in the cultured and characterized human small intestinal lamina propria fibroblasts in relation to neutrophil migration and adhesion in response to lipopolysaccharide (LPS) extracted from EPEC 055:B5 . Upon interaction with LPS (1-10 micrograms/ml), lamina propria fibroblasts produced a high level of proinflammatory mediators, interleukin (IL)-1alpha, IL-1beta, IL-6, IL-8, tumor necrosis factor (TNF)-alpha and cell adhesion molecules (CAM) such as intercellular cell adhesion molecule (ICAM), A-CAM, N-CAM and vitronectin in a time-dependent manner . LPS induced cell-associated IL-1alpha and IL-1beta, and IL-6, IL-8 and TNF-alpha as soluble form in the supernatant . Apart from ICAM, vitronectin, A-CAM, and N-CAM proteins were strongly induced in lamina propria fibroblasts by LPS . Adhesion of PBMC to LPS-treated lamina propria fibroblasts was ICAM-dependent . LPS-induced ICAM expression in lamina propria fibroblasts was modulated by whole blood, PBMC and neutrophils . Conditioned medium of LPS-treated lamina propria fibroblasts remarkably enhanced the neutrophil migration . The migration of neutrophils was inhibited by anti-IL-8 antibody . Co-culture of fibroblasts with neutrophils using polycarbonate membrane filters exhibited time-dependent migration of neutrophils . These findings indicate that the coordinate production of proinflammatory cytokines and adhesion molecules in lamina propria fibroblasts which do not classically belong to the immune system can influence the local inflammatory reactions at the intestinal mucosal site during bacterial infections and can influence the immune cell population residing in the lamina propria. J Gastroenterol Hepatol, 1999 Feb, 14(2), 146 - 9 Findings and benefit of liver biopsies in 46 patients infected with human immunodeficiency virus; Piratvisuth T et al.; AIMS: The aim of this work is to evaluate the role of liver biopsy and to determine the histological findings in patients infected with the human immunodeficiency virus (HIV) who have abnormal liver function tests (LFT) . METHODS: We performed a percutaneous liver biopsy in 46 HIV-seropositive patients with abnormal LFT . Parts of biopsied tissue were used for bacterial and fungal culture and the rest was processed for histological examination including special staining . RESULTS: Of these 46 patients, 41 patients were males and five were females . The median age was 31+/-6 years . Mycobacterium tuberculosis was the most common histological finding (15 cases) . Of 15 tuberculosis patients, 11 (73.3%) had lymphadenopathy and positive acid-fast bacilli (AFB) in node aspiration or biopsy . The other findings included AFB-negative granuloma (eight cases), histoplasmosis (six cases), cryptococcosis (six cases), penicillosis (four cases), viral hepatitis: hepatitis C virus (HCV; one case), hepatitis B virus and HCV infection (one case), fatty liver (two cases), drug-induced hepatitis (one case) and non-specific changes (five cases) . There were double infections in three patients . We were able to demonstrate opportunistic infections in 41 cases (89.3%) . CONCLUSIONS: Mycobacterium tuberculosis was the most common histological finding in HIV patients with abnormal LFT in Thailand . Liver biopsy was a useful procedure in evaluating abnormal LFT in HIV patients. Clin Infect Dis, 1999 Jan, 28(1), 82 - 92 Early mycological treatment failure in AIDS-associated cryptococcal meningitis; Robinson PA et al.; Cryptococcal meningitis causes significant morbidity and mortality in persons with AIDS . Of 236 AIDS patients treated with amphotericin B plus flucytosine, 29 (12%) died within 2 weeks and 62 (26%) died before 10 weeks . Just 129 (55%) of 236 patients were alive with negative cerebrospinal fluid (CSF) cultures at 10 weeks . Multivariate analyses identified that titer of cryptococcal antigen in CSF, serum albumin level, and CD4 cell count, together with dose of amphotericin B, had the strongest joint association with failure to achieve negative CSF cultures by day 14 . Among patients with similar CSF cryptococcal antigen titers, CD4 cell counts, and serum albumin levels, the odds of failure at week 10 for those without negative CSF cultures by day 14 was five times that for those with negative CSF cultures by day 14 (odds ratio, 5.0; 95% confidence interval, 2.2-10.9) . Prognosis is dismal for patients with AIDS-related cryptococcal meningitis . Multivariate analyses identified three components that, along with initial treatment, have the strongest joint association with early outcome . Clearly, more effective initial therapy and patient management strategies that address immune function and nutritional status are needed to improve outcomes of this disease. Am J Kidney Dis, 1999 Feb, 33(2), 282 - 6 Renal pathology and HIV infection in Thailand; Praditpornsilpa K et al.; The existence of a human immunodeficiency virus (HIV)-associated nephropathy (HIVAN) as a distinct disease entity characterized by glomerulosclerosis is well established in North America and Western Europe . Although the large number of HIV-infected cases overwhelm the Asian countries, no cases of HIVAN are documented in the literature . We studied 26 cases of HIV-infected Thai patients with proteinuria greater than 1.5 g/d of protein during 1995 and 1996 . None of the patients were treated with antiretroviral drugs at the time of renal biopsy . Intravenous drug addiction and sexual transmission were risk factors in 11 and 15 patients, respectively . Pathological examinations were performed by light microscopic and immunoperoxidase study . Mesangial proliferative glomerulonephritis was found in 17 cases, immunoglobulin A (IgA) nephropathy in 2 cases, and diffuse proliferative glomerulonephritis and interstitial nephritis secondary to cryptococcal infection in 2 cases each . One case each had membranous glomerulopathy, membranoproliferative glomerulonephritis, and granulomatous interstitial nephritis secondary to tuberculosis . The renal pathological findings of HIVAN with the unique features described in previous literature were not evident in these patients . Although the data in this study are limited to 26 HIV-infected Thai patients, we believe that HIVAN is uncommon in the Asian HIV-infected population. Gene, 1999 Feb 18, 227(2), 231 - 40 The yeast Cryptococcus neoformans uses 'mammalian' enhancer sites in the regulation of the virulence gene, CNLAC1; Zhang S et al.; Transcriptional regulation in mammalian and plant cells is distinguished from fungi by the presence of blocks of multiple interacting DNA binding sites distributed over a relatively large upstream region of genes and the ability to use glutamine-rich enhancers such as Sp1 . We offer evidence that the haploid yeast Cryptococcus neoformans contains a virulence gene, CNLAC1, having regulatory properties more similar to mammalian systems than to that of yeast . We used a novel promoter plasmid, pVEW, and electromobility shift assay techniques adapted for the fungus for the first systematic structural and functional study of a 5'-enhancer region of a basidiomycete fungus using the upstream region of CNLAC1 . Two groups of interactive enhancer regions, located over a range of 1.5kb from the mRNA start site are involved in CNLAC1 regulation (region 2: -1721 to -1615 and region 7) in addition to a TATA promoter at position -539 . Region 2 contains a consensus Sp1 site and region 7 contains a consensus E2F site, each of which shows significant binding to nuclear proteins under derepressed conditions; cooperative binding was also suggested between DNA-binding protein of these sites and those binding nearby CCAAT sequences in each region . Two regions of repression were also evident under derepressed conditions (region 5: -1351 to -1207 and region 8: -991 to -971) . Identification of functional Sp1 binding sites and the presence of multiple interactive enhancer sites over a fairly large upstream range suggests that cryptococcal transcriptional regulation contains features often associated with higher eukaryotic regulation . C . neoformans thus may provide a unique system for the study of certain aspects of higher eukaryotic transcription, using yeast genetic approaches . In addition, properties of basidiomycete yeast such as Cryptococcus exemplified in the present study suggest an evolutionary progression in gene regulation within fungi toward properties exhibited in the kingdoms Animalia and Plantae. Med Mycol, 1998, 36 Suppl 1, 57 - 67 Development of vaccines and their use in the prevention of fungal infections; Dixon DM et al.; Vaccine approaches to infectious diseases are widely applied and appreciated . Disciplines such as bacteriology and virology have a rich history of successful vaccine development . The complexity of eukaryotic systems presents additional challenges to the development of vaccines against them . These challenges are being met in the fields of parasitology, and are being revisited for application in oncology . Vaccine opportunities exist in medical mycology . The National Institute of Allergy and Infectious Diseases has held a series of workshops in medical mycology where the need to develop vaccines for fungal diseases was noted and where important opportunities were discussed . Major advances in vaccinology and the technology of antigen preparation and delivery have increased feasibility and heightened interest . The recent epidemic of coccidioidomycosis in the American Southwest has demonstrated the need for developing a vaccine as an effective preventive measure for those living in and for those who subsequently move into regions with the endemic mycoses . The XIIth Congress of the International Society for Human and Animal Mycology included a symposium that summarized new vaccination strategies for selected fungi: Candida albicans, Coccidioides immitis, and Trichophyton verrucosum . The goal of the present summary is to provide representative examples of continuing efforts relating to vaccine development within the medical mycological community highlighting Blastomyces dermatidis, Cryptococcus neoformans, Histoplasma capsulatum, Paracoccidioides brasiliensis, and Pythiumn insidiosum. Med Mycol, 1998, 36 Suppl 1, 249 - 57 Molecular diagnosis and epidemiology of fungal infections; Reiss E et al.; A variety of methods are utilized for DNA strain subtyping of Candida spp . because no 'gold standard' exists . Random amplified polymorphic DNA (RAPD) or restriction enzyme analysis (REA) are useful to determine the source of an outbreak, but more reproducible and discriminatory methods such as Southern hybridization and pulsed field gel electrophoresis (PFGE) may be required . When applied to some nosocomial Candida infections, multiple strains and species have been identified . Microevolution of yeast species occurs and epidemiologically related isolates may show minor pattern differences, creating uncertainty as to whether they are distinct strains . Approximately 1000 isolates of Aspergillus fumigatus from environmental and clinical sources were typed by REA probed with an A . fumigatus-specific retrotransposon-like sequence . Patients with no symptom of aspergillosis may carry several strains, whereas patients with pulmonary aspergillosis may carry one or two strains; nocosomial transmission of aspergillosis was proven in 39% of the patients studied; any given environmental strain can be infectious; the environmental population of A . fumigatus is extremely diverse and no specific niche was found in the hospital . A PCR assay was designed to target conserved 18S-ribosomal DNA (rDNA) sequences shared by most fungi and a 687 bp product was amplified from 25 medically important fungal species . Studies with blood, cerebrospinal fluid and sputum specimens from patients with mycoses indicated that the PCR assay is more sensitive in diagnosing invasive fungal infections than blood culture methods . More specific identification is obtainable with genus/species-specif c probes designed from within the PCR-amplified sequences for C . albicans, C . krusei, C . lusitaniae, Pneumocystis carinii, Cryptococcus neoformans, Aspergillus/Penicillium spp . and C . glabrata/Saccharomyces cerevisiae . A . fumigatus and A . niger were differentiated by denaturing gradient gel electrophoresis . In situ hybridization (ISH) detected a 648 bp fragment of the 18S rDNA of C . neoformans and a 568 bp fragment of the alkaline proteinase gene of A . fumigatus in tissues from experimentally infected animals . In ISH, the entire process can be automated, making this procedure rapid and easy . The difficulty in establishing a diagnosis of invasive candidiasis has prompted the quest for a clinically useful PCR test for candidaemia . The universal fungal oligonucleotide primer pair, ITS3 and ITS4, amplifies portions of the 5.8S ad 28S rDNA subunits, and the ITS2 region . Although rRNA genes are highly conserved, the ITS regions are distinctive . DNA probes were designed from ITS2 that were specific for 16 different Candida species . Simple, rapid sample preparation was suitable for PCR analysis of BacT/Alert blood culture bottles . Sample preparation, PCR, and EIA detection of the amplicon from five different Candida species was accomplished in 7 h, 2.5 days sooner than by conventional culture methods . As well as saving time, minor yeast species among a major species, or among bacteria, were simultaneously detected . PCR-EIA using a microtitration plate format had sensitivity 10-times greater than that obtained with ethidium bromide-stained agarose gels . Taqman combines in one step PCR, probe hybridization, and fluorescent signal generation . Taqman PCR had sensitivity equivalent to PCR-EIA and required only 5 h, including sample preparation. Med Mycol, 1998, 36 Suppl 1, 207 - 15 Recent advances in cryptococcosis, candidiasis and coccidioidomycosis complicating HIV infection; Kappe R et al.; Concomitant with the decline in CD4+ T-cells seen as human immunodeficiency virus (HIV) infection progresses, the prevalence of opportunistic mycoses increases dramatically . This article reviews selected recent advances in our understanding of the immunology, molecular epidemiology and treatment of fungal infections in patients infected with HIV . For cryptococcosis, studies are reported on how HIV infection affects the immune response to Cryptococcus neoformans and, conversely, how stimulation with C . neoformans induces HIV production from latently HIV-infected cells . In addition, studies are presented examining the efficacy of triple combination antimycotic chemotherapy in cryptococcosis . For candidosis, investigations into genetic profiles of Candida albicans isolates obtained from patients, with resistance to antifungal agents, are demonstrated . Finally, for coccidioidomycosis, prospective studies are presented examining the clinical, epidemiological and immunological characteristics of a cohort of HIV-infected subjects residing in an endemic area. Med Mycol, 1998, 36 Suppl 1, 119 - 28 Antifungal drug resistance in pathogenic fungi; Vanden Bossche H et al.; Failures of drug treatment in fungal infections combined with improvements in performances and standardization of antifungal susceptibility testing have drawn attention to the problem of antifungal resistance and its underlying mechanisms . Resistance of Candida species and Cryptococcus neoformans to flucytosine (5FC) develops during monotherapy . Acquired resistance results from a failure to metabolize 5FC to 5FUTP and 5FdUMP, or from the loss of feedback control of pyrimidine biosynthesis . A combination of 5FC and amphotericin B (AmB) reduces the appearance of resistant C . albicans isolates . Resistance to AmB is unusual . C . lusitaniae is the most susceptible to AmB resistance . C . neoformans with decreased AmB susceptibility has been isolated from an HIV-infected patient . Acquired resistance to AmB is often associated with alteration of membrane lipids, especially ergosterol . Concomitant with the widespread use of fluconazole there have been increasing reports of fluconazole resistance in Candida species and C . neoformans . Fluconazole resistance was mostly associated with prior use of fluconazole as intermittent therapy or prophylactic continuous treatment for recurrent thrush . In contrast to fluconazole, itraconazole is active against C . krusei . Decreased susceptibility to itraconazole is observed over time in C . albicans isolates becoming resistant to fluconazole . Decreased susceptibility to itraconazole and SCH-56592 was also observed in a few Aspergillus fumigatus isolates . Failure to accumulate azole antifungals has been identified as a cause of resistance in several post-treatment C . albicans, C . glabrata and C . krusei isolates . In azole-resistant C . albicans isolates from AIDS patients with oropharyngeal candidiasis, multidrug efflux transporters of the ATP-binding cassette (ABC) superfamily and of the class of major facilitators (MF) have been shown to be responsible for the low level of accumulation of azole antifungal agents . Two genes for these transporters, the ABC-transporter gene CDR1 and the MF gene, CaMDR1 (BEN) were shown to be overexpressed in resistant C . albicans isolates . Overexpression of BEN in Saccharomyces cerevisiae conferred resistance to fluconazole and terbinafine . CDR1 overexpression in S . cerevisiae conferred cross-resistance to fluconazole, itraconazole, ketoconazole and terbinafine . C . albicans clinical isolates resistant to azole antifungal agents over-expressing the ABC-transporter genes CDR1 and CDR2 were less susceptible to the morpholine derivative amorolfine . In C . glabrata isolates azole resistance is based on over-expression of the CgCDR gene . A reduced susceptibility of ergosterol biosynthesis is another mechanism of resistance described in a number of post-treatment C . albicans, C . neoformans and Histoplasma capsulatum isolates . Mutations have been reported in the CYP51A1 genes of resistant C . albicans isolates . Over-expression of CYP51A1 in C . albicans and C . glabrata may also account for a decreased susceptibility to azole antifungal agents. Med Mycol, 1998, 36 Suppl 1, 79 - 86 Cryptococcus neoformans: virulence and host defences; Perfect JR et al.; Cryptococcus neoformans represents a model organism for the study of virulence and the host response . In this discussion, there is a focus on the genetic, molecular, and biochemical aspects of C . neoformans as it interacts with the host . Investigations into direct and indirect virulence phenotypes are now possible . The molecular aspects of two major virulence factors, capsule and melanin, are characterized . Yeast polyol metabolism through mannitol is examined as a potential biochemical pathway for virulence . The concept of C . neoformans differentially expressed genes within the host or in response to certain environmental cues can be used indirectly to identify potential virulence genes . However, despite significant progress in molecular pathogenesis with C . neoformans, the future of research in this area will require a certain critical mass of investigators to help share in the developmental costs which continue to occur. Med Mycol, 1998, 36 Suppl 1, 38 - 44 Fate of transforming DNA in pathogenic fungi; Kwon-Chung KJ et al.; Genetic engineering is an important tool in helping us to define the molecular basis of pathogenicity and is also useful in helping us to identify new therapeutic targets in pathogenic fungi . Molecular genetic manipulation of micro-organisms requires the development of plasmid-mediated transformation systems that include: (i) infusion of exogenous DNA into recipient cells, (ii) expression of genes present on the incoming DNA, and (iii) stable maintenance and replication of the inserted DNA leading to expression of the desired phenotypic trait . Transformation systems have been developed for only a handful of fungi that are pathogenic to humans including several species of Candida, Cryptococcus neoformans, Histoplasma capsulatum, Blastomyces derrmatitidis, Aspergillus fumigatus, Wangiella dermatitidis (Exophiala dermatitidis) and Coccidioides immitis . Except for Candida species and A . fumigatus, where passage of exogenous DNA into recipient cells has been achieved readily using methods developed for transformation of Saccharomyces cerevisiae and Aspergillus nidulans, respectively, development of transformation systems in other pathogenic fungi has been delayed considerably and has only been possible recently with the introduction of electroporation and biolistic methods . Conventional spheroplasting methods or cell wall permeabilization methods using lithium acetate have not been successful for transformation of C . neoformans and work with only low efficiency in H . capsulatum . The fate of incoming DNA varies greatly in these pathogenic species regardless of their phylogenetic relationships . Understanding the fate of incoming DNA is critical for the construction of transforming vectors and the molecular manipulation of the organisms . In this symposium, recent advances in molecular genetic systems including transformation systems, the fate of incoming DNA and strategies for targeted integration are discussed in relation to four pathogenic fungi. Mayo Clin Proc, 1999 Jan, 74(1), 78 - 100 Antifungal agents . Part II . The azoles; Terrell CL; Before 1978, amphotericin B and flucytosine were the only drugs available for the treatment of systemic fungal infections . The imidazoles, miconazole and ketoconazole, were introduced during the next 3 years . Intravenously administered miconazole served a limited therapeutic role and is no longer available . Orally administered ketoconazole, an inexpensive, effective, and convenient option for treating mucosal candidiasis, was widely used for a decade because it was the only available oral therapy for systemic fungal infections . During the 1990s, use of ketoconazole diminished because of the release of the triazoles--fluconazole and itraconazole . Fluconazole is less toxic and has several pharmacologic advantages over ketoconazole, including penetration into the cerebrospinal fluid . In addition, it has superior efficacy against systemic candidiasis, cryptococcosis, and coccidioidomycosis . Despite a myriad of drug interactions and less favorable pharmacologic and toxicity profiles in comparison with fluconazole, itraconazole has become a valuable addition to the antifungal armamentarium . It has excellent activity against sporotrichosis and seems promising in the treatment of aspergillosis . Itraconazole has replaced ketoconazole as the therapy of choice for nonmeningeal, non-life-threatening cases of histoplasmosis, blastomycosis, and paracoccidioidomycosis and is effective in patients with cryptococcosis and coccidioidomycosis, including those with meningitis . Further investigation into the development of new antifungal agents is ongoing. J Clin Microbiol, 1999 Mar, 37(3), 838 - 40 Cryptococcus neoformans var . grubii: separate varietal status for Cryptococcus neoformans serotype A isolates; Franzot SP et al.; Cryptococcus neoformans var . neoformans presently includes isolates which have been determined by the immunologic reactivity of their capsular polysaccharides to be serotype A and those which have been determined to be serotype D . However, recent analyses of the URA5 sequences and DNA fingerprinting patterns suggest significant genetic differences between the two serotypes . Therefore, we propose to recognize these genotypic distinctions, as well as previously reported phenotypic differences, by restricting C . neoformans var . neoformans to isolates which are serotype D and describing a new variety, C . neoformans var . grubii, for serotype A isolates. J Clin Microbiol, 1999 Mar, 37(3), 715 - 20 Genetic multilocus studies of different strains of Cryptococcus neoformans: taxonomy and genetic structure; Bertout S et al.; The genotypes of 107 strains of Cryptococcus isolated from the environment or from patients from various geographical areas were determined by multilocus enzyme electrophoresis (MLEE) . We analyzed the relationships between genotype structure and serotype and between genotype structure and strain origin . Twelve of the 14 enzyme-encoding loci studied were polymorphic, giving rise to 48 electrophoretic types . The genotypes of C . neoformans and C . laurentii were very similar . MLEE could not distinguish between these two pathogenic species . A correlation between the genetic multilocus structure and the origin of the sample (from the environment or patients) existed . A second analysis detected a correlation between genotype distribution and serotype . The second analysis considered three serotype groups (B, C, and A plus D plus A/D), proving that serotypes A, D, and A/D are closely related . MLEE is a useful epidemiological tool for improving our understanding of the biology of this fungus. J Clin Microbiol, 1999 Mar, 37(3), 694 - 9 PCR detection of DNA specific for Trichosporon species in serum of patients with disseminated trichosporonosis; Nagai H et al.; Deep-seated trichosporonosis is a lethal opportunistic infection that disseminates rapidly and widely in immunocompromised patients, and early diagnosis is crucial for the treatment of this infection . We developed a novel nested-PCR assay that detects DNA specific for clinically important strains of Trichosporon in serum samples from patients with disseminated trichosporonosis . In this assay, two sets of oligonucleotide primers were derived from the sequence of 26S rRNA genes of Trichosporon asahii . The specific fragment was amplified from T . asahii and T . mucoides, but not from other microorganisms, including some other basidiomycetous fungi (Cryptococcus, Malassezia, Rhodotorula, and Sporobolomyces) . Target DNA was detected by the nested PCR with as little as 5 fg of the extracted DNA of T . asahii . In a study using 11 clinical samples, the specific fragment was detected by the nested PCR in 64% (7 of 11) of sera from patients with histologically diagnosed disseminated trichosporonosis, while glucuronoxylomannan antigen was detected in only 54% (6 of 11) of the samples . Our new nested-PCR assay using serum samples can be performed repeatedly throughout the course of the disease . In addition, not only can it be used for early diagnosis of trichosporonosis, but it may also be beneficial for monitoring its progress or response to therapy. J Clin Microbiol, 1999 Mar, 37(3), 591 - 5 Multicenter comparison of the sensititre YeastOne Colorimetric Antifungal Panel with the National Committee for Clinical Laboratory standards M27-A reference method for testing clinical isolates of common and emerging Candida spp., Cryptococcus spp., and other yeasts and yeast-like organisms; Espinel-Ingroff A et al.; National Committee for Clinical Laboratory Standards (NCCLS) standard guidelines are available for the antifungal susceptibility testing of common Candida spp . and Cryptococcus neoformans, but NCCLS methods may not be the most efficient and convenient procedures for use in the clinical laboratory . MICs of amphotericin B, fluconazole, flucytosine, itraconazole, and ketoconazole were determined by the commercially prepared Sensititre YeastOne Colorimetric Antifungal Panel and by the NCCLS M27-A broth microdilution method for 1,176 clinical isolates of yeasts and yeast-like organisms, including Blastoschizomyces capitatus, Cryptococcus spp., 14 common and emerging species of Candida, Hansenula anomala, Rhodotorula spp., Saccharomyces cerevisiae, Sporobolomyces salmonicolor, and Trichosporon beigelii . Colorimetric MICs of amphotericin B corresponded to the first blue well (no growth), and MICs of the other agents corresponded to the first purple or blue well . Three comparisons of MIC pairs by the two methods were evaluated to obtain percentages of agreement: 24- and 48-h MICs and 24-h colorimetric versus 48-h reference MICs . The best performance of the YeastOne panel was with 24-h MICs (92 to 100%) with the azoles and flucytosine for all the species tested, with the exception of C . albicans (87 to 90%) . For amphotericin B, the best agreement between the methods was with 48-h MIC pairs (92 to 99%) for most of the species tested . The exception was for isolates of C . neoformans (76%) . These data suggest the potential value of the YeastOne panel for use in the clinical laboratory. J Immunol, 1999 Feb 1, 162(3), 1618 - 23 Specific activated T cells regulate IL-12 production by human monocytes stimulated with Cryptococcus neoformans; Retini C et al.; IL-12 production mediated by a T cell-independent and/or T cell-dependent pathway was investigated in human monocytes responding to Cryptococcus neoformans . The data of this study showed that: 1) appreciable levels of IL-12 were observed when freshly isolated monocytes were exposed to acapsular C . neoformans or Candida albicans and secretion occurred within 24-48 h of incubation; 2) monocytes alone were poor producers of IL-12 when stimulated with encapsulated C . neoformans; 3) the presence of specific anti-glucuronoxylomannan mAb favored IL-12 secretion and Fc cross-linking could play a role; 4) monocytes were able to secrete consistent levels of IL-12 when cultured with activated T cells responding to C . neoformans; 5) the maximum secretion of IL-12 was observed at 5-7 days of culture and was strongly regulated by the presence of endogenous IFN-gamma; and 6) the interaction between CD40 on monocytes and CD40 ligand on activated T lymphocytes responding to C . neoformans played a critical role in IL-12 secretion . These data highlight the mechanisms of IL-12 production by human monocytes exposed to C . neoformans, indicating a possible biphasic secretion of IL-12, dependent on the direct effect of fungal insult, and characterized by consistent secretion of IL-12 that is dependent on the interaction of CD40 with the CD40 ligand expressed on activated T cells responding to C . neoformans. Recenti Prog Med, 1998 Dec, 89(12), 657 - 67 {AIDS and opportunistic visceral mycoses . A diagnostic and therapeutic update}; Manfredi R; Candidiasis, cryptococcosis, aspergillosis, histoplasmosis, and coccidioidomycosis are the most commonly recognized visceral mycoses complicating the course of HIV infection and AIDS . Their current management issues are discussed on the ground of the personal experience, and the most recent literature evidences. Nippon Ishinkin Gakkai Zasshi, 1999, 40(1), 31 - 4 Ploidy of serotype AD strains of Cryptococcus neoformans; Tanaka R et al.; Twenty-four serotype AD strains of Cryptococcus neoformans were tested for ploidy and mating type . These included 13 natural isolates (1 from a patient, 1 from peach juice, 11 from pigeon droppings), 2 single clones from one of these natural isolates, 7 F1 progeny from a self-fertile strain and 2 F1 progeny of a cross between a strain from a patient (serotype A, alfa-mating type: MATalfa) and a tester strain (serotype D, a-mating type: MATa) . Six strains (2 of natural isolates, 1 of single clone and 3 of F1 progeny) were MATalfa, 1 of F1 progeny was MATa, 4 (1 of natural isolate, 1 of single clone and 2 of F1 progeny) were a-alfa-mating type (MATa-alfa) and 13 (10 of natural isolates and 3 of F1 progeny) were untypable . Most strains, regardless of mating type, were diploid. Nippon Ishinkin Gakkai Zasshi, 1999, 40(1), 9 - 14 {Dermatomycosis in human and animals}; Nakamura Y et al.; Dermatomycosis including dermatophytosis, sporotrichosis and cryptococcosis commonly occurs in humans and animals all and are considered to be zoonotic diseases . Recently, human cases of dermatophytosis transmitted from animals are increasing in number due to changes in the environments of human and animal life . Three species of dermatophytes, Microsporum canis, Trichophyton mentagrophytes and T . verrucosum are the most important pathogens from animal to human, respectively . Therefore, it is necessary to understand their biological and ecological characteristics to correctly diagnose and treat the disease . Some human cases of sporotrichosis and cryptococcosis were reported to be transmitted from animals in Europe and America, suggesting that medical doctors should be careful in taking the history of human patients with reference to their contacts with animals . Close cooperation between medical and veterinary doctors is required in clinical studies on mycotic infection. Mycopathologia, 1998, 142(2), 71 - 6 Comparison of phospholipase production in Cryptococcus neoformans isolates from AIDS patients and bird droppings; Vidotto V et al.; Secreted phospholipase has been recently proposed as a virulence determinant in Cryptococcus neoformans as well as Candida albicans . This issue of cryptococcal phospholipase requires screening of phospholipase production in a larger number of isolates from clinical and environmental sources . In this study we examined phospholipase production in a total of 67 C . neoformans isolates from AIDS patients and bird droppings by using the egg-yolk plate method . Phenoloxidase activity, capsule size and growth at 37 degrees C were also measured in these strains in order to observe a possible relationship between phospholipase production of different C . neoformans strains and its virulence . Four of the 21 AIDS strains at 28 degrees C and 1 at 37 degrees C did not produce phospholipase, respectively . In contrast, 38 and 34 of the 46 bird dropping strains were negative for phospholipase production at 28, and 37 degrees C, respectively . Statistical analysis revealed a significant difference in phospholipase production, capsule size and growth ability at 37 degrees C, but not phenoloxidase activity, between the AIDS and the bird dropping strains . The highly prevalent distribution of phospholipase activity in the AIDS strains suggests a role of the enzyme in invading the host. Antimicrob Agents Chemother, 1999 Feb, 43(2), 413 - 4 An alternative animal model for comparison of treatments for cryptococcal meningitis; Najvar LK et al.; Weanling outbred rats were infected with Cryptococcus neoformans by direct percranial puncture and inoculation into the cranium . A lethal infection ensued . Treatment with LY295337, a depsipeptide with antifungal activity, was effective in prolonging survival and reducing fungal counts in brain tissue . Weanling rats are an acceptable model for the study of central nervous system infection with C . neoformans. Antimicrob Agents Chemother, 1999 Feb, 43(2), 233 - 9 Amphotericin B and fluconazole affect cellular charge, macrophage phagocytosis, and cellular morphology of Cryptococcus neoformans at subinhibitory concentrations; Nosanchuk JD et al.; Amphotericin B (AmB) and fluconazole (FLU) are the major antifungal drugs used in the treatment of cryptococcosis . Both drugs are believed to exert their antifungal effects through actions on cell membrane sterols . In this study we investigated whether AmB and FLU had other, more subtle effects on C . neoformans that could contribute to their therapeutic efficacy . C . neoformans cells were grown in media with subinhibitory concentrations of either AmB or FLU and analyzed for cellular charge, phagocytosis by macrophages with antibody and complement opsonins, appearance by scanning electron and light microscopies, and release of the capsular polysaccharide glucuronoxylomannan into the culture medium . Growth in the presence of either AmB or FLU resulted in major reductions in cellular charge, as measured by determination of the zeta potential . Phagocytosis studies demonstrated that exposure of C . neoformans to subinhibitory concentrations of AmB or FLU enhanced phagocytosis by macrophages . Scanning electron microscopy revealed that a large proportion of cells had an altered capsular appearance . Cells grown in medium with either AmB or FLU were smaller and released more glucuronoxylomannan into the culture medium than cells grown without antibiotics . The results suggest additional mechanisms of action for AmB and FLU that may be operative in body compartments where drug levels do not achieve the MICs . Furthermore, the results suggest mechanisms by which AmB and FLU can cooperate with humoral and cellular immune defense systems in controlling C . neoformans infections. Med Clin (Barc), 1998 Dec 5, 111(19), 725 - 30 {Pulmonary cavitation lesions in patients infected with the human immunodeficiency virus: an analysis of a series of 78 cases}; Rodriguez Arrondo F et al.; BACKGROUND: To assess the clinical, radiologic and microbiological features of lung cavitation and HIV infection . Evaluation of the differences related to this disease in the last years . PATIENTS AND METHODS: Retrospective review of all patients with lung cavitation and HIV infection admitted at our hospital from January 1989 until December 1994 and prospective study of all patients with the same characteristics during 1995 and 1996 . Lung cavitation was defined as any parenchymal lesion, with air content, visible in a simple X-ray and greater than 1 cm of diameter . Criteria for confirmed, probable or possible diagnosis were defined . RESULTS: 78 cases of lung cavitation have been identified in 73 patients . The radiologic patterns included unilobar and multilobular involvement in 31 and 47 cases, respectively . Cavities were multiple and single in 40 and 38 cases respectively . Findings with fine needle aspiration biopsy (FNAB) were diagnostic in 11 out of 14 cases . A clinical diagnosis was performed in all 78 cases, with microbiological results in 69 cases (88.5%): Mycobacterium tuberculosis in 20, Pneumocystis carinii in nine, Pseudomonas aeruginosa in nine, Staphylococcus aureus in eight (5 endocarditis with cavitary septic emboli), Rhodococcus equi in six, P . aeruginosa and S . aureus in three, Salmonella enteritidis in three, Cryptococcus neoformans in two, Aspergillus fumigatus in two and others in 7 cases . Confirmed, probable and possible diagnosis was considered in 54, 15 and 9 cases, respectively . Thirteen episodes of spontaneous pneumothorax were found . CONCLUSIONS: The lung cavitation rate is low, compared with the number of admissions related to HIV infection; nevertheless, many of them are in close relationship with HIV infection, and most of them are caused by treatable infections . It is important to know the clinical and radiological characteristics, in order to establish an early diagnosis and an appropriate therapy . Pseudomonas aeruginosa is becoming an important cause of lung cavitation . In our series, spontaneous pneumo-thorax was not related to Pneumocystis carinii pneumonia in 61.5% of cases. Rev Hosp Clin Fac Med Sao Paulo, 1998 Jul-Aug, 53(4), 184 - 8 {Immunohistochemistry diagnosis of fungal infections}; Rocha DC et al.; Because the mycosis incidence has increased a lot with the appearing of AIDS, the Immunohistochemistry study among fungus shows the importance of fast methods for their identification that have advantage of been a durable method in comparison with immunofluorescence and the possibility of making retrospective studies in material embedded in paraffin . The Immunohistochemistry reaction with Histoplasma capsulatum, Pneumocystis carinii; and Criptococcus neoformans antibodies were sensitive, specifics, and intensely positive in all the cases previously diagnosed as Histoplasmosis, Pneumocystosis and Cryptococcosis, without cross-reaction with other fungus; while the anti-Candida albicans antibody showed weak positiveness in four Histoplasmosis cases, in one of Paracoccidioidomycosis cases and Sporotrichosis case; and the reactions with the antibody anti-P . brasiliensis were intensely positive in all the Paracoccidioidomycosis cases and weakly positive in two Histoplasmosis and two of the four Candidiasis cases . The previous identification of each fungi on tissue sample was made by Grocott method . This preliminary study showed that it is necessary to use other kinds of antibody and fungus, in order to get more details about the possible occurrence of cross-reactions . We suggest the use of new antibodies, with new standardizations in order to find the best titles for each reaction and eliminate the cross-reactions. J Thorac Imaging, 1999 Jan, 14(1), 51 - 62 Opportunistic fungal pneumonia; Connolly JE Jr et al.; Opportunistic fungal infection is a common cause of serious morbidity and mortality in immunocompromised patients . These infections occur primarily in patients with chemotherapy-induced neutropenia, acquired immunodeficiency syndrome . or immunosuppression after solid organ or bone marrow transplantation . The most important opportunistic fungal pathogens include Cryptococcus neoformans, Candida and Aspergillus species, and the fungi that cause mucormycosis . Opportunistic pneumonia caused by previously unrecognized pathogens, such as Fusarium, Penicillium, and the dematiaceous fungi, are increasingly reported . The clinical and radiologic features of opportunistic fungal pneumonia are highly variable and often nonspecific . Diagnosis requires knowledge of the various modes of presentation, radiologic manifestations, and epidemiology of these infections . Because many of these organisms can colonize the upper airway, sputum cultures are considered diagnostically unreliable . Instead, definitive diagnosis requires culture of the fungus from infected tissue or demonstration of the organism on microscopic examination. Am J Pathol, 1999 Jan, 154(1), 145 - 52 Expression of inducible nitric oxide synthase in human granulomas and histiocytic reactions; Facchetti F et al.; Inducible nitric oxide synthase (iNOS) is required in immune response against infections and is involved in granuloma formation in animals; in murine macrophages, iNOS is induced by lipopolysaccharide and interferon-gamma . In contrast, the role of iNOS in human immune response against infections is still questioned, and its expression in granulomas is poorly investigated . Using Western blotting and immunohistochemistry, we investigated iNOS expression in human lymph nodes with nonspecific reactions and in tissues containing granulomas caused by mycobacteria, Toxoplasma, Cryptococcus neoformans, Leishmania, Bartonella, noninfectious granulomas (sarcoidosis, foreign body), and other hystiocitic reactions (Kikuchi's disease, Omenn syndrome) . iNOS was undetectable in nonspecific reactive lymphadenitis, foreign-body granulomas, and Omenn syndrome, whereas it was strongly expressed in infectious granulomas, sarcoidosis, and Kikuchi's diseases . Immunohistochemistry demonstrated that iNOS was selectively expressed by the epithelioid and multinucleated giant cells within the granulomas . Use of an anti-nitrotyrosine antibody, recognizing nitrosilated amino acid residues derived from nitric oxide production, revealed a consistent positivity within the cells expressing iNOS, thus suggesting that iNOS is functionally active . Detection of cytokines by reverse transcriptase-polymerase chain reaction demonstrated that tissues that were positive for iNOS, also expressed the Thl-type cytokine interferon-gamma mRNA, but not the Th2-type cytokine interleukin-4 . Taken together, these results indicate that iNOS is involved in different human immune reactions characterized by histiocytic/granulomatous inflammation and associated with Th1-type cytokine secretion. Mycoses, 1998 Nov, 41(9-10), 389 - 96 Phenotypic and genotypic differentiation of several human and avian isolates of Cryptococcus neoformans; Hotzel H et al.; The Cryptococcus neoformans strains isolated from two human cases could be diagnosed as Cr . neoformans var . neoformans by differentiation on the basis of their characteristics determined by proline, canavanine and EDTA urease tests . The results of the serovar assignment were: for the isolate from the meningoencephalitis patient with lethal outcome, serovar A; for the strain isolated from the osteomyelitis patient with benign course, serovar D . Also, the PCR fingerprinting using primers (GACA)4, (CAC)5 and FM 1 resulted in a clear and reproducible assignment of the Cr . neoformans strains to the varieties neoformans and gattii, respectively, and, in addition, it confirmed the serovar assignment . No statistically confirmed differences in virulence between the osteomyelitis and the meningoencephalitis strain could be established by i.v . testing in mice, nor did the PCR with several primers provide any clues to a genetically determined higher virulence of the meningoencephalitis strain . The different classification as serovars A and D does not allow any conclusions concerning different virulence . It was not possible to retrospectively establish the sources of infection of the two Cr . neoformans infections, but pigeon faeces may well have played a role as a reservoir for one of the illnesses. Kansenshogaku Zasshi, 1998 Dec, 72(12), 1261 - 8 {Clinical study on fluconazole (FLCZ) in the treatment of primary pulmonary cryptococcosis}; Hayashi Y et al.; We treated two cases of primary pulmonary cryptococcosis with fluconazole (FLCZ), the clinical usefulness of FLCZ was evaluated . FLCZ was administered orally in doses of 300 mg daily for about six months . Concentrations of FLCZ were measured in the serum of the two cases and in the bronchoalveolar lavage (BAL) fluid in one case . The following results were obtained: 1 . Clinical cures were obtained in the two cases . 2 . The serum levels of FLCZ was 15.1 microliters/ml, 13.6 micrograms/ml two hours after administration of 100 mg in case 1, that of levels were 11.1 micrograms/ml, 8.9 micrograms/ml one hour and 4.5 hours, respectively, after administration of 100 mg in case 2 . BAL was performed 4.5 hours after administration of 100 mg in case 2, the BAL fluid level of FLCZ was 0.7 microgram/ml . 3 . The minimal inhibitory concentration of FLCZ against one strain obtained from the cytology brush in case 1 was 4.0 micrograms/ml . 4 . The cryptococcal antigen titer decreased with the improvement of clinical signs and the resolution of chest X-ray abnormalities within about six months, and there was no relapse . From these results, we consider that FLCZ is a useful antifungal agent for primary pulmonary cryptococcosis, and we therefore recommend a six month treatment. Infect Immun, 1999 Feb, 67(2), 936 - 41 The cell wall and membrane of Cryptococcus neoformans possess a mitogen for human T lymphocytes; Mody CH et al.; The mechanism of human T-lymphocyte activation by the pathogenic yeast Cryptococcus neoformans has not been established . Previous investigations have suggested that C . neoformans contains a mitogen for T lymphocytes, while other investigators have attributed lymphocyte proliferation in vitro to a recall antigen . Because of the potential importance of the mechanism of T-cell activation for our understanding of the immune response to C . neoformans, the present studies were performed to determine whether C . neoformans contains a mitogen for T lymphocytes . C . neoformans stimulates fetal blood lymphocytes to proliferate and stimulates proliferation of CD45RA+ cells from adults, indicating that it stimulates naive T cells . The T-cell response to C . neoformans was dependent upon the presence of accessory cells . However, allogeneic cells were sufficient for accessory cell function, indicating that the response was not major histocompatibility complex restricted . The percentage of T cells in the cell cycle was higher than that with the recall antigen tetanus toxoid but lower than that with the mitogenic lectin phytohemagglutinin A or the superantigen Staphylococcus enterotoxin B . Precursor frequency analysis established that 1 in 7,750 +/- 2, 270 T cells proliferated in response to the cryptococcal cell wall and membrane . Compared to the case for most mitogens or superantigens, the proliferative response is late and the number of T cells that enter the cell cycle and the precursor frequency are low, indicating that the mitogenic effect is modest . However, the mitogenic effect of C . neoformans should be considered when interpreting the immune response to C . neoformans, since even weak mitogens can have profound effects on host defense. Infect Immun, 1999 Feb, 67(2), 885 - 90 Cryptococcus neoformans resides in an acidic phagolysosome of human macrophages; Levitz SM et al.; Recently, we demonstrated that human monocyte-derived macrophages (MDM) treated with chloroquine or ammonium chloride had markedly increased antifungal activity against the AIDS-related pathogen Cryptococcus neoformans . Both of these agents raise the lysosomal pH, which suggested that the increased antifungal activity was a function of alkalinizing the phagolysosome . Moreover, there was an inverse correlation between growth of C . neoformans in cell-free media and pH . These data suggested that C . neoformans was well adapted to survive within acidic compartments . To test this hypothesis, we performed studies to determine the pH of human MDM and neutrophil phagosomes containing C . neoformans . Fungi were labeled with the isothiocyanate derivatives of two pH-sensitive probes: fluorescein and 2',7'-difluorofluorescein (Oregon Green) . These probes have pKas of 6.4 and 4.7, respectively, allowing sensitive pH detection over a broad range . The phagosomal pH averaged approximately 5 after ingestion of either live or heat-killed fungi and remained relatively constant over time, which suggested that C . neoformans does not actively regulate the pH of its phagosome . The addition of 10 and 100 microM chloroquine resulted in increases in the phagosomal pH from a baseline of 5.1 up to 6.5 and 7.3, respectively . Finally, by immunofluorescence, colocalization of C . neoformans and the MDM lysosomal membrane protein LAMP-1 was demonstrated, establishing that fusion of C . neoformans-laden phagosomes with lysosomal compartments takes place . Thus, unlike many other intracellular pathogens, C . neoformans does not avoid fusion with macrophage lysosomal compartments but rather resides and survives in an acidic phagolysosome. Nihon Kokyuki Gakkai Zasshi, 1998 Oct, 36(10), 902 - 7 {Spontaneous resolution of pulmonary cryptococcosis--report of 2 cases}; Kakeya H et al.; We encountered two patients with pulmonary cryptococcosis that resolved spontaneously without antifungal treatment . A 59-year-old man (patient 1) and 37-year-old man (patient 2) were admitted to our hospital for the examination of abnormal shadows on their chest x-ray films . Transbronchial lung biopsies were performed, and cryptococci were detected in both patient's lung tissues . Latex aggultination tests for cryptococcal antigen were positive (1:16 and 1:4, respectively) . The size of the shadows reduced spontaneously and the titers of the cryptococcal antigen in sera decreased within three months although no antifungal therapy was performed . Bronchoalveolar lavage fluid (BALF) was obtained from each patient and examined . In patient 1, the lymphocyte to CD 4/8 ratio was high in the BALF obtained on admission and the lymphocyte count was low; however, the CD 4/8 ratio was still high in a BALF sample obtained 3 months after discharge . In patient 2, the findings of the BALF analysis on admission were almost normal . The analysis of BALF may contribute to the immunological conditions to in the patients with pulmonary cryptococcosis. J Clin Microbiol, 1999 Feb, 37(2), 315 - 20 New PCR primer pairs specific for Cryptococcus neoformans serotype A or B prepared on the basis of random amplified polymorphic DNA fingerprint pattern analyses; Aoki FH et al.; Thirty-three strains of Cryptococcus neoformans were isolated from clinical specimens, including specimens from AIDS patients in Brazil, and were classified into two serotypes; we detected 31 and 2 strains of serotypes A and B, respectively . Random amplified polymorphic DNA (RAPD) fingerprint pattern analyses of these strains of serotypes A and B showed that the patterns were similar for strains of each serotype when three 10-mer primers were used as the RAPD primers . Comparative studies of the fingerprint patterns of the study isolates with those of the reference strains also showed that the RAPD patterns for strains of each serotype were related and that most of the fingerprint bands existed commonly for all strains of each serotype tested . The common RAPD bands (an approximately 700-bp band for serotype A and an approximately 450-bp band for serotype B) were extracted and the DNA sequences were determined . Using this information, we prepared two and one PCR primer pairs which were expected to be specific for C . neoformans serotypes A and B, respectively . Use of each PCR primer combination thus prepared for serotype A or B was 100% successful in identifying the respective C . neoformans serotypes, including the 33 clinical isolates tested in the present study . Among these combinations, one for serotype A was found to amplify DNA from C . neoformans serotype B as well as serotype A . Serotype B-specific PCR primer pairs amplified DNA from not only serotype B strains but also from serotype C strains . The usefulness of other serotype-specific PCR primers for clinical C . neoformans isolates is discussed. Diagn Microbiol Infect Dis, 1998 Nov, 32(3), 223 - 7 Evaluation of Etest for determining in vitro susceptibility of yeast isolates to amphotericin B; Pfaller MA et al.; We evaluated the performance of Etest using several different agar media for testing of amphotericin B against 660 clinical isolates of yeast species including Candida albicans, C . glabrata, C . tropicalis, C . parapsilosis, C . lusitaniae, C . krusei, Candida spp., Cryptococcus neoformans, and Saccharomyces cerevisiae . Two of the C . lusitaniae isolates represented strains with high-level amphotericin B resistance . All isolates were tested by NCCLS microdilution methods with RPMI 1640 medium and by Etest using RPMI agar with 2% glucose (RPG) . A subset of 108 isolates was also tested by Etest using RPG, antibiotic medium 3 agar (AM3), Casitone agar (CAS), and Mueller-Hinton agar (MHA) . The overall agreement between the NCCLS reference method and Etest using RPG was 98.3% . All of the Etest methods identified the two resistant strains (MICs, 4.0 to 16 micrograms/mL), whereas the reference method failed to distinguish them from 18 other isolates with MICs of 2.0 micrograms/mL . Among the 20 isolates with reference MICs of 2.0 micrograms/mL, 12 had MICs > or = 2.0 micrograms/mL when tested by Etest with RPG (range 2.0 to 16 micrograms/mL) compared with eight with AM3, two with CAS, and five with MHA . These data indicate that Etest identifies subpopulations of yeast isolates with high amphotericin B MICs . The greater sensitivity of Etest for detection of amphotericin B resistance should be exploited in future surveillance studies. Diagn Microbiol Infect Dis, 1998 Nov, 32(3), 191 - 9 Molecular epidemiology and antifungal susceptibility of Cryptococcus neoformans isolates from Ugandan AIDS patients; Pfaller M et al.; Little is known of the antifungal susceptibility patterns and molecular epidemiology of Cryptococcus neoformans from tropical regions . We studied 164 clinical isolates of C . neofomans from 120 Ugandan AIDS patients with cryptococcal meningitis by analyzing their electrophoretic karyotypes and antifungal susceptibility profiles . Computer-assisted analysis of karyotype patterns was performed to generate dendrograms . MICs of fluconazole and flucytosine were determined by reference methods . A total of 43 distinguishable DNA types were identified among the 164 isolates . Only 30 patients (25%) were infected with their own unique strain of c . neoformans, whereas 75% of the patients shared their infecting strain with at least one other patient . Among 17 patients with more than one CSF isolate of C . neoformans, sequential isolates were identical or highly related in 12 (71%) and were different in five patients (29%) . The isolates were susceptible to both fluconazole and flucytosine and there were no instances in which a stepwise increase in either fluconazole or flucytosine MICs was observed among serial isolates . These findings suggest that the epidemiology of cryptococcal disease in AIDS patients from tropical regions may be somewhat different from that observed in more temperate climatesPIP: Even though Cryptococcus neoformans var . neoformans is a leading cause of life-threatening mycotic infection among AIDS patients worldwide, little is known about its antifungal susceptibility patterns and molecular epidemiology in tropical regions . The authors studied 164 clinical isolates of C . neoformans from 120 Ugandan AIDS patients with cryptococcal meningitis by analyzing their electrophoretic karyotypes and antifungal susceptibility profiles . Computer-assisted analysis of karyotype patterns was performed to generate dendrograms, while the MICs of fluconazole and flucytosine were determined using reference methods . 43 distinguishable C . neoformans DNA types were identified among the 164 isolates . 30 patients (25%) were infected with their own unique strain of C . neoformans, while 75% of the patients shared their infecting strain with at least 1 other patient . Among 17 patients with more than 1 cerebrospinal fluid isolate of C . neoformans, sequential isolates were identical or highly related in 12 (71%) and were different in 5 patients (29%) . The isolates were susceptible to both fluconazole and flucytosine, and there was no instance in which a stepwise increase in either fluconazole or flucytosine MICs was observed among serial isolates . These findings suggest that the epidemiology of cryptococcal disease in AIDS patients from tropical regions may be somewhat different from that observed in more temperate climates . J Bacteriol, 1999 Jan, 181(2), 444 - 53 A glucan synthase FKS1 homolog in cryptococcus neoformans is single copy and encodes an essential function; Thompson JR et al.; Cryptococcal meningitis is a fungal infection, caused by Cryptococcus neoformans, which is prevalent in immunocompromised patient populations . Treatment failures of this disease are emerging in the clinic, usually associated with long-term treatment with existing antifungal agents . The fungal cell wall is an attractive target for drug therapy because the syntheses of cell wall glucan and chitin are processes that are absent in mammalian cells . Echinocandins comprise a class of lipopeptide compounds known to inhibit 1,3-beta-glucan synthesis, and at least two compounds belonging to this class are currently in clinical trials as therapy for life-threatening fungal infections . Studies of Saccharomyces cerevisiae and Candida albicans mutants identify the membrane-spanning subunit of glucan synthase, encoded by the FKS genes, as the molecular target of echinocandins . In vitro, the echinocandins show potent antifungal activity against Candida and Aspergillus species but are much less potent against C . neoformans . In order to examine why C . neoformans cells are less susceptible to echinocandin treatment, we have cloned a homolog of S . cerevisiae FKS1 from C . neoformans . We have developed a generalized method to evaluate the essentiality of genes in Cryptococcus and applied it to the FKS1 gene . The method relies on homologous integrative transformation with a plasmid that can integrate in two orientations, only one of which will disrupt the target gene function . The results of this analysis suggest that the C . neoformans FKS1 gene is essential for viability . The C . neoformans FKS1 sequence is closely related to the FKS1 sequences from other fungal species and appears to be single copy in C . neoformans . Furthermore, amino acid residues known to be critical for echinocandin susceptibility in Saccharomyces are conserved in the C . neoformans FKS1 sequence. Clin Microbiol Rev, 1999 Jan, 12(1), 126 - 46 The evolutionary biology and population genetics underlying fungal strain typing; Taylor JW et al.; Strain typing of medically important fungi and fungal population genetics have been stimulated by new methods of tapping DNA variation . The aim of this contribution is to show how awareness of fungal population genetics can increase the utility of strain typing to better serve the interests of medical mycology . Knowing two basic features of fungal population biology, the mode of reproduction and genetic differentiation or isolation, can give medical mycologists information about the intraspecific groups that are worth identifying and the number and type of markers that would be needed to do so . The same evolutionary information can be just as valuable for the selection of fungi for development and testing of pharmaceuticals or vaccines . The many methods of analyzing DNA variation are evaluated in light of the need for polymorphic loci that are well characterized, simple, independent, and stable . Traditional population genetic and new phylogenetic methods for analyzing mode of reproduction, genetic differentiation, and isolation are reviewed . Strain typing and population genetic reports are examined for six medically important species: Coccidioides immitis, Histoplasma capsulatum, Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, and A . flavus . Research opportunities in the areas of genomics, correlation of clinical variation with genetic variation, amount of recombination, and standardization of approach are suggested. J Infect Dis, 1999 Feb, 179(2), 449 - 54 Cryptococcosis: population-based multistate active surveillance and risk factors in human immunodeficiency virus-infected persons . Cryptococcal Active Surveillance Group; Hajjeh RA et al.; To determine the incidence of cryptococcosis and its risk factors among human immunodeficiency virus (HIV)-infected persons, population-based active surveillance was conducted in four US areas (population, 12.5 million) during 1992-1994, and a case-control study was done . Of 1083 cases, 931 (86%) occurred in HIV-infected persons . The annual incidence of cryptococcosis per 1000 among persons living with AIDS ranged from 17 (San Francisco, 1994) to 66 (Atlanta, 1992) and decreased significantly in these cities during 1992-1994 . Among non-HIV-infected persons, the annual incidence of cryptococcosis ranged from 0.2 to 0.9/100,000 . Multivariate analysis of the case-control study (158 cases and 423 controls) revealed smoking and outdoor occupations to be significantly associated with an increased risk of cryptococcosis; receiving fluconazole within 3 months before enrollment was associated with a decreased risk for cryptococcosis . Further studies are needed to better describe persons with AIDS currently developing cryptococcosis in the era of highly active antiretroviral therapy. Am J Respir Cell Mol Biol, 1999 Jan, 20(1), 135 - 42 Phenotypic and functional characterization of normal rat pleural macrophages in comparison with autologous peritoneal and alveolar macrophages; Gjomarkaj M et al.; Pleural mononuclear phagocytes (PleMP) were isolated from normal rats by pleural lavage and compared with autologous peritoneal (PerMP) and bronchoalveolar mononuclear phagocytes (BAMP) isolated by peritoneal and bronchoalveolar lavage, respectively . The phagocytic activity of PleMP, PerMP, and BAMP, evaluated by testing their ability to ingest latex beads, was lower for PleMP and PerMP than for BAMP . The phenotype of PleMP, PerMP, and BAMP was characterized by immunocytochemical staining with a panel of monoclonal antibodies (mAbs) . As expected, PleMP, PerMP, and BAMP did not react with OX19, OX33, ED5, MOM/3F12/F2, and anticytokeratin mAbs, specific for T lymphocytes, B lymphocytes, dendritic cells, granulocytes, and epithelial/mesothelial cells, respectively . Moreover, PleMP and PerMP populations were highly enriched with OX6-, OX42-, ED7-, and ED8-positive MP, whereas BAMP population was enriched with ED1- and ED9-positive cells . To test the ability of PleMP, PerMP, and BAMP to function as accessory cells (AC), mitomycin C-treated MP were used as stimulatory cells in mixed leukocyte reaction experiments, using allogeneic T cells as responders . 3HdTR incorporation by T cells was assessed as an index of AC function . PleMP and PerMP were more potent AC than BAMP . Moreover, when cultured together with autologous pulmonary interstitial dendritic cells, PleMP and PerMP exerted a more potent ability to stimulate T-cell proliferation than did BAMP . To investigate the capacity of MP to function as bactericidal and fungicidal cells, we tested their ability to kill Escherichia coli and Cryptococcus neoformans, respectively . PleMP and PerMP were less potent bactericidal and fungicidal cells than BAMP . The results of this study demonstrate that PleMP isolated from normal rat pleural space are functionally and phenotypically different from BAMP but similar to PerMP, and suggest that these cells might play an important role in cell-mediated immune reactions in the pleural space. Arch Pathol Lab Med, 1998 Dec, 122(12), 1107 - 11 Fontana-Masson--stained tissue from culture-proven mycoses; Kimura M et al.; BACKGROUND: The Fontana-Masson stain can be used to detect melanin of dematiaceous fungi such as Curvularia lunata and Bipolaris hawaiiensis in tissue . This stain is often used to identify fungi that may not have an evident brown color in hematoxylin-eosin-stained sections . However, it is not clear whether hyphae of nondematiaceous fungi can be stained with the Fontana-Masson procedure . METHODS: Paraffin sections from culture-proven mycoses caused by species of Aspergillus, Candida, Fusarium, Pseudallescheria, Trichosporon, zygomycetes, and dematiaceous fungi were stained by the Fontana-Masson procedure . Extent of stain intensity and its distribution in fungal elements in tissue were evaluated by means of an intensity scale based on Cryptococcus neoformans as a positive control . RESULTS: Many Aspergillus spp, some zygomycetes, and all of the dematiaceous fungi showed strong intensity, whereas other fungi showed either weak intensity or no staining . CONCLUSION: Fungal hyphae other than dematiaceous fungi occasionally can be stained with the Fontana-Masson procedure . The morphology of positively stained hyphae must be carefully evaluated, especially when they are hyaline in hematoxylin-eosin-stained tissue sections. Antimicrob Agents Chemother, 1999 Jan, 43(1), 169 - 71 In vitro activities of voriconazole, fluconazole, and itraconazole against 566 clinical isolates of Cryptococcus neoformans from the United States and Africa; Pfaller MA et al.; We investigated the in vitro activity of voriconazole compared to those of fluconazole and itraconazole against 566 clinical isolates of Cryptococcus neoformans from Africa (164) and the United States (402) . Isolates were obtained from cerebrospinal fluid (362), blood (139), and miscellaneous sites (65) . Voriconazole (MIC at which 90% of the isolates are inhibited {MIC90}, 0.12 to 0.25 microg/ml) was more active than either itraconazole (MIC90, 0.5 microg/ml) or fluconazole (MIC90, 8.0 to 16 microg/ml) against both African and U . S . isolates . Isolates inhibited by >/=16 microg of fluconazole per ml were almost all (99%) inhibited by </=1 microg of voriconazole per ml . These results suggest that voriconazole may be useful in the treatment of cryptococcosis. Antimicrob Agents Chemother, 1999 Jan, 43(1), 106 - 14 Mixture-based heterocyclic combinatorial positional scanning libraries: discovery of bicyclic guanidines having potent antifungal activities against Candida albicans and Cryptococcus neoformans; Blondelle SE et al.; A mixture-based synthetic combinatorial library of more than 100,000 bicyclic guanidines was generated in a positional scanning format and assayed for activity against Candida albicans . Potent individual bicyclic guanidines were directly identified following the screening of the library . Time-kill curve studies indicated bactericidal activities for the individual bicyclic guanidines . These compounds also showed potent activity against Cryptococcus neoformans . These studies demonstrate the value of using mixture-based combinatorial positional scanning libraries made up of heterocyclic compounds for the rapid identification of novel classes of antifungal compounds. Arch Pharm Res, 1998 Dec, 21(6), 688 - 91 Growth inhibitory activities of kalopanaxsaponins A and I against human pathogenic fungi; Kim DW et al.; Antifungal activities of the compounds isolated from Kalopanax pictus against representative fungi of dermatomycosis were investigated using paper disc diffusion method . It was found that kalopanaxsaponins A and I were effective in inhibiting the growth of Candida albicans KCTC 1940 and Cryptococcus neoformans KCTC 7224 with minimum inhibitory concentration (MIC) of 25 micrograms/ml . It showed that antifungal activity of both compounds have strong selectivity against the fungi of dermatomycosis. Mayo Clin Proc, 1998 Dec, 73(12), 1205 - 25 Antifungal agents . Part I . Amphotericin B preparations and flucytosine; Patel R; Traditionally, amphotericin B has been the cornerstone of antifungal treatment . Toxicity, however, is a major dose-limiting factor of amphotericin B deoxycholate . Nevertheless, it continues to have a major role in the treatment of deep-seated mycotic infections . Recently, less nephrotic lipid formulations, including amphotericin B lipid complex, amphotericin B cholesteryl sulfate, and liposomal amphotericin B, have been introduced . The pharmacologic properties, main indications, recommended dosages, related costs, and adverse effects of these various preparations are summarized in this review . Orally administered flucytosine is useful in certain infections, particularly cryptococcal meningitis, but it should be used with caution in patients with renal insufficiency. Infect Immun, 1999 Jan, 67(1), 220 - 9 Mechanisms for induction of L-selectin loss from T lymphocytes by a cryptococcal polysaccharide, glucuronoxylomannan; Dong ZM et al.; Disseminated cryptococcosis is accompanied by cryptococcal polysaccharides in the serum and the lack of cellular infiltrates in infected tissues . Cryptococcal polysaccharides given intravenously to mice inhibit the influx of T lymphocytes into the sites of cell-mediated immune response . The focus here was to determine whether cryptococcal polysaccharides modulate the expression of molecules, such as L-selectin, that are important in extravasation of T cells . Cryptococcal glucuronoxylomannan (GXM), but not galactoxylomannan or mannoprotein, was found to cause loss of L-selectin from freshly isolated human T cells of both CD4 and CD8 subsets and from Jurkat cells . With the signaling-pathway inhibitors staurosporine (which inhibits protein kinase C) and herbimycin A (which inhibits protein tyrosine kinases), we showed that GXM or the cryptococcal culture filtrate antigen CneF directly induces L-selectin loss from CD4(+) and CD8(+) T cells via a herbimycin A-sensitive pathway(s) presumably involving one or more protein tyrosine kinases but not via a pathway involving protein kinase C . Loss of L-selectin from the T cells before the T cells have a chance to bind to L-selectin ligands on endothelial cells would be expected to prevent T-cell migration into inflamed tissues and/or lymph organs. Infect Immun, 1999 Jan, 67(1), 108 - 12 Catecholamine oxidative products, but not melanin, are produced by Cryptococcus neoformans during neuropathogenesis in mice; Liu L et al.; Melanin has been proposed as a virulence factor in Cryptococcus neoformans, but its presence has not been shown unambiguously in vivo . Validated methods used previously to show production of cryptococcal eumelanin pigment in vitro (P . R . Williamson, K . Wakamatsu, and S . Ito, J . Bacteriol . 180:1570-1572, 1998) were used to assess for production of laccase-derived products in mouse brain of the Lacc+ strains, 2E-TUC, H99 (serotype A), and ATCC 34873 (serotype D), and the Lacc- strain, 2E-TU . Pyrrole-2,3, 5-tricarboxylic and pyrrole-2,3-dicarboxylic acid, specific degradation products of catecholamine derivatives such as melanin, were found in all Lacc+ strains, but not in the Lacc- strain, 2E-TU . However, the presence of melanin pigment itself could not be demonstrated in the same cells . Lack of the specific degradation products aminohydroxyphenylalanine and aminohydroxyphenylethylamine in Lacc+ strains upon hydriodic acid hydrolysis showed that pheomelanin was also not produced by the fungus in vivo . These are the first data to support the generation of catecholamine oxidation products by C . neoformans in vivo, but they do not support postenzymatic polymerization of these products to form typical eumelanin, as previously proposed. Wei Sheng Wu Xue Bao, 1997 Apr, 37(2), 135 - 41 {Polymorphism and pologenetic affinities of genotype in pathogenic yeast}; Li D et al.; The classification of pathogenic yeast, which is one of the most common reasons of opportunistic infection in human body, will be useful for the epidemiological survey and clinical works . The genotypic polymorphism of intergenus, interspecies and intraspecies in 48 isolates, including Candida, Cryptococcus, Torulopsis, Trichosporon, Saccaromyces had been administered carefully with patterns amplified by RAPD (Randomly amplified polymorphic DNA) in 53 primers . The pologenetic affinities had been evaluated by similarity coefficients obtained from these profiles . The results indicated that there are significant difference among intergenus, interspecies and intraspecies in Candida and related yeast . The similarity coefficients among Candida and Crytococcus, Trichosporon were maintained about 80% . The similarity coefficients among interspecies of Candida ranged from 82%-87%, and ones of intraspecies of different Candida species were more than 90% . The genotypic typing of species except C . guilliermondii seems to be related to morphological classification. Mycoses, 1998 Sep-Oct, 41(7-8), 277 - 80 Fungaemia due to Cryptococcus laurentii and a review of non-neoformans cryptococcaemia; Johnson LB et al.; Cryptococcus laurentii is one of several non-neoformans cryptococci that have rarely been associated with human infection . The spectrum of clinical infection due to non-neoformans species ranges from skin lesions to fungaemia . Most cases of non-neoformans fungaemia have been nosocomially acquired and have been associated with indwelling intravascular catheters and neutropenia . Limited data on in vitro susceptibilities of non-neoformans cryptococci show these species to be more resistant to fluconazole and flucytosine than most Cr . neoformans . Two such cases are presented here. Enferm Infecc Microbiol Clin, 1998, 16 Suppl 1, 29 - 35 {Infections by Candida and cryptococci}; Berenguer J et al.; The introduction of highly active antiretroviral therapy (HAART) has reduced dramatically the incidence of mucosal candidiasis and cryptococcosis in AIDS patients . Fluconazole is the drug of choice for candidiasis . The duration of antifungal treatment is based on response, but typically 7 to 14 days are required for oropharyngeal forms and up to 21 days for esophageal disease (200 the first day and 100 mg thereafter) . Resistant candidiasis tends to occur in persons with advanced HIV disease and previous fluconazole therapy who have been noted to result in clinical improvement with HAART . HAART must be considered the therapy of choice for refractory candidiasis . The preferred treatment for cryptococcal meningitis includes two weeks induction treatment with amphotericin B (0.7 mg/kg/d IV) with or without flucytosine (25 mg/kg qid) followed by 8 weeks of fluconazole (200-400 mg PO qd) . Long-term maintenance therapy with fluconazole (200-400 mg PO qd) is required to prevent relapses . In patients with elevated intracranial pressure who had focal neurologic deficits or mental status changes, serial lumbar punctures should be performed . In refractory cases the immediate placement of CSF drains must be considered. Mol Cell Biol, 1999 Jan, 19(1), 745 - 50 Melanization of Cryptococcus neoformans in murine infection; Nosanchuk JD et al.; Cryptococcus neoformans is a fungus that is pathogenic in humans and that can produce melanin in vitro . Melanization is associated with virulence, but there is no evidence that melanin is made during infection . Melanins are difficult to study because they are amorphous and insoluble . Melanin-binding peptides from a phage display library were used to demonstrate that C . neoformans makes melanin-like compounds in tissue . Melanin-binding peptides were characterized by a high proportion of positively charged and aromatic residues .